Real-time RT-PCR for Mayaro virus detection in plasma and urine

Autor: Jesse J. Waggoner, Benjamin A. Pinsky, Yvalena Guillén, Alejandra Rojas, Alisha Mohamed-Hadley
Rok vydání: 2018
Předmět:
Zdroj: Journal of Clinical Virology. 98:1-4
ISSN: 1386-6532
DOI: 10.1016/j.jcv.2017.11.006
Popis: Background Mayaro virus (MAYV) causes an acute febrile illness which can be difficult to differentiate from dengue or chikungunya. MAYV RNA can be detected in plasma during the first 3–5 days of illness, but only a single rRT-PCR has been fully evaluated in the literature. Objectives To develop an rRT-PCR for MAYV and evaluate assay performance using human plasma and urine samples spiked with different MAYV strains. Study design A MAYV rRT-PCR targeting a region of the 5′UTR and nsp1 gene was designed from the alignment of all complete-genome MAYV sequences to be compatible with existing laboratory protocols. The assay was evaluated using human samples spiked with six MAYV strains, including strains from each of the three genotypes. Results The linear range of the MAYV rRT-PCR extended from 1.0 to 8.0 log10 copies/μL, and the lower limit of 95% detection was 8.2 copies/μL. No detection was observed when the MAYV rRT-PCR was tested with genomic RNA from related arboviruses. The assay demonstrated linear amplification of all 6 MAYV strains when spiked into human plasma samples as well as 2 strains spiked into urine. Conclusions We report the design and evaluation of an rRT-PCR for MAYV. Given the concern for MAYV emergence in the Americas and the few molecular tests that have been evaluated in the literature, this assay should provide a useful diagnostic for patients with an acute febrile illness.
Databáze: OpenAIRE
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